Pharmaceutical compositions comprising 72-methyl estrone and methods for using same

ABSTRACT

D R A W I N G

United States Patent Primary Examiner-Elbert L. RobertsAttorneys-Willard L. Cheesman and John Kekich ABSTRACT: This inventionrelates to novel steroid ticularly to those compounds embraced by theformula (ll) 7 compounds and processes for their preparation; more par-RO "CH3 wherein R is selected from the group consisting of hydrogen, theacyl radical of a hydrocarbon carboxylic acid containing from onethrough twelve carbon atoms, an alkyl radical containing from onethrough eight carbon atoms, tetrahydror a a x ltqt a lslway nY ubwi le lyd ap y andasilylrad icaloithe formula 72] Inventors John C. Babcock;

J. Allan Campbell, both of Kalamazoo, Mich. [2]] Appl. No. 666,488 [22]Filed Sept. 8, 1967 [45] Patented Dec. 14, 1971 [73] Assignee The UpjohnCompany Kalamazoo, Mich. Continuation-impart of application Ser. No.114,621, June 5, 1961, now Patent No. 3,341,557, Continuation-impart ofapplication Ser. No. 69,557, Nov. 6, 1960, now abandoned. Thisapplication Sept. 8, I967, Ser. No. 666,488

[54] PHARMACEUTICAL COMPOSITIONS COMPRISING 72-METIIYL ESTRONE ANDMETHODS FOR USING SAME 4 Claims, No Drawings [52] US. Cl 424/243,260/239.55,260/397.4, 195/51 51 im. CI 'c07c 169/20; [50] Field ofSearch /I-Iand $35 M S99E12 [5 6] References Cited UNITED STATES PATENTS3,318,925 5/1967 Anner et al. 260/3975 FOREIGN PATENTS 2/1966 France swherein R1, R and K, are selected from the group consisting of alkyl ofone through six carbon atoms and phenyl.

PHARMACEUTICAL COMPOSITIONS COMPRISING 72- METI'IYL ESTRONE AND METHODSFOR USING SAME CROSS-REFERENCES TO RELATED APPLICATIONS This applicationis a continuation-in-part of Application Ser. No. 114,621, filed June 5,1961, now Pat. No. 3,341,557 which is in turn a continuation-in-part ofabandoned Application Ser. No. 69,557, filed Nov. 6, 1960.

BRIEF SUMMARY OF THE INVENTION 7a-methylestrone, embraced by formula 11,above, can be prepared by several known methods described below.

1. By fermentation of 7a-methyl-l9-nortestosterone (l) or7a-methyl-l9-nor-4-androstene-3,l7-dione with a micro-organism or itsenzymes capable of introducing a double bond in the 1(2) or 1(2)- and4(5)-positions of the steroid nucleus, e.g., Corynebacterium simplex orSeptomyxa affim's to yield 70:- methylestrone l l).

2. By catalytically dehydrogenating 7a-methyll9-nor-4-androstene-3,l7-dione at the 1(2)-positions, e.g., by heating inthe presence of a hydrogenation catalyst (e.g., palladium on charcoal)in a high boiling solvent (e.g., cymene), to give7amethylestrone l l 3.By treating 7a-methyl-l9-nor-4-androstene3,l7-dione with a chemicaldehydrogenating agent, e.g., a quinone such as2,3-dichloro-5,6-dicyanobenzoquinone (DDQ) or 2,3,5,6-tetrachloro-l,4-benzoquinone (chloranil), or selenium compounds such asselenium dioxide or dibenzoyloxy selenium oxide, to yield7a-methylestrone (l l).

4. By pyrolysis of 7a-methyl-l,4-androstadiene-3,l7-dione at elevatedtemperatures (e.g., between about 400 to 600 C.) in high boilingdiluents (e.g., heavy mineral oil) to give 70:- methylestrone (l l).

5. By treating 7a-methyl-l,4-androstadiene-3,17-dione in accordance withthe procedures described in J. Amer. Chem. Soc. 86, 742, i.e., withlithium and diphenyl in the presence of diphenylmethane and employingtetrahydrofuran as solvent, to yield 7a-methylestrone (l 1).

The 3-ethers of 7a-methylestrone of formula 11, above, can be preparedby the known methods described below.

1. By treating 7a-methylestrone (11) in accordance with the proceduresdisclosed in British Pat. No. 909,662, i.e., with an alkyl (orcycloalkyl) halide and an alkali metal alkylate, preferably at refluxtemperature, to give a 3-alkyl (or cycloalkyl) ether of 7a-methylestrone(l l). 1

2. By treating 7a-methylestrone (11) with an alkylating agent (e.g., adialkylsulfate) in conventional manner, to give a 3-alkyl ether of7a-methylestrone (l l 3. By treating 7a-methylestrone (11) with a cyclicenol ether (e.g., dihydrofuran, dihydropyran,S-hydroxymethyldihydropyran, S-carboxydihydropyran, etc.) at lowtemperature, preferably in the presence of an acidic catalyst (e.g.,phosphorus oxychloride), to give the corresponding 3-ether (e.g.,tetrahydrofuranyl, tetrahydropyranyl, S-hydroxymethyltetrahydropyranyl,etc.) of 7a-methylestrone l l).

4. By treating 7a-methylestrone (l l) with a diazoalkane (e.g.,diazomethane, diazoethane, diazobutane, etc.) at ambient temperature inan inert solvent such as ether, ethylene glycol dimethyl ether, etc., togive the corresponding 3-alkyl ether of 7a-methylestrone l l 5. Bytreating 7a-methylestrone (l l) with a disilazane of the fonnula NHwherein R R, and R, have the same meaning as above (e.g.,hexamethyldisilazane, symmetrical diphenyltetramethyldisilazane,l-methyll l -dibutyl-3-phenyl-3,3- dirnethyldisilazane,hexa-amyldisilazane, etc.) to yield a corresponding 3-silyl ether of7a-methylestrone l l The 3-acylates of 7a-methylestrone of formula 11,above, can be prepared by conventional procedures, e.g., by treating7a-methylestrone l l) with the appropriate organic carboxylic acidanhydride (or chloride) at moderate temperatures in the presence of anesterification catalyst such as pyridine. to yield the corresponding7a-methylestrone 3-acylate l l All of the compounds included withinformula 11, above, can be isolated from their respective reactionmixtures by conventional means, for example,'when a water-misciblesolvent is used, by pouring the reaction mixture into water andseparating the resulting precipitate by filtration or by extraction withwater-immiscible solvents. Additional purification of the products canbe accomplished by conventional means, for example, by elutionchromatography from an adsorbent column with a suitable solvent such asacetone, methanol, dilute methanol, ethanol, ether, methylene chlorideand Skellysolve B (hexanes), mixtures and combinations of thesesolvents; also by gradient elution chromatography from an adsorbentcolumn with a suitable mixture of solvents, such as methylenechloride-Skellysolve B, acetone-Skellysolve B, and the like, and bycrystallization. v

The compounds of formula of the present invention are potent estrogens'when administered either orally or parenterally; e.g., when assayedin'rats by the Allen-Doisy test 7a-methylestrone l l was found to haveapproximatelythree times the activity of estrone. These compounds havealso demonstrated marked antifertility activity in the male and femalerat. These compounds when administered to mammals, birds and animals,are also active in lowering cholesterol in the blood, inhibitinggonadatropin secretion, producing anabolic response, especially inproviding nitrogen retention, and in supplying calcium lost as a resultof osteoporosis. In addition, the compounds of formula 1 1, whencombined with progestins such as fia-methyl-17a-hydroxyprogesteronel7-acetate (Provera), 7a-methyll 7o4-ethylnyll 9-nortestosterone, 17ahydroxy-6-methyll 6-methylene-4,6-pregnadiene-3 ,20-dione l7-acetate(Melengestrol acetate), l7-hydroxyl 9-nor-l 7apregn-5(l0)-en-20-yn-3-one (Norethynodrel), l9-nor-l 7apregn-4-en-20yne-3fi,l7-diol 3,17-diacetate (Ethynodiol diacetate),l7-hydroxy-l9-nor-l7a-pregn-4 en-20-yn-3-one (Norethindrone), thecorresponding l7-acetate (Norethindrone acetate),601,21-dimethyl-l7B-hydroxy-4-pregnen-20- yn-3-one (Dimethisterone),6-chlorol 7a-hydroxy-4,6- pregnadiene-3,20-dione l7-acetate, etc., areuseful for the prevention of ovulation in mammals. The foregoingproperties make the new compounds useful in veterinary practice.

The compounds of the invention can be prepared and administered tomammals, birds, and animals, in the wide variety of oral or parenteraldosage forms, singly or in admixture with other coacting compounds. Theycan be administered with a pharmaceutical carrier which can be a solidmaterial or a liquid in which the compound is dissolved, dispersed orsuspended. The solid compositions can take the form of tablets, powders,capsules, pills, or the like, preferably in unit dosage forms for simpleadministration or precise dosages. The liquid compositions can take theform of solutions, emulsions, suspensions, syrups, or elixirs.

DETAILED DESCRlPTlON It is to be understood that the invention is not tobe limited to the exact details of operation or exact compositions shownand described herein, as obvious modifications and equivalents will beapparent to one skilled in the art, and the invention is therefore to belimited only by the scope of the appended claims.

EXAMPLE I 7a-methylestrone( 7a-methyll ,3 ,5( 10 )-estratrien-3-oll 7-one) (1 l) To 10 l. of a sterile glucose-yeast extract medium there wasadded an inoculum of 500 ml. of Corynebacterium simplex ATCC 6946. Theculture was stirred and aerated for about 48 hours at a temperature ofabout 28 C., lard oil being added to suppress the foam. After about 48hours the pH was 6.1. To

the fermentor, l g. of 7a-methyl-l9-nortestosterone (1), prepared as inSteroids l, 317, was added and aeration continued for about'24 hours. Atthis time the pH was 6.5. The beer was adjusted to pH 3 withhydrochloric acid and extracted four times with 3 l. of methylenechloride. Paper chromatography of an aliquot of the solvent extractindicated, by the Bush 13-3 system, that essentially all of the7a-methyl-l9- nortestosterone (1) had been reacted and that twovcompounds showing the characteristics of aromatic A ring steroids werepresent. The less polar compound had the mobility of l. 7a-methylestrone(11) and the more polar that of 70:- methylestradiol. The methylenechloride extracts were evaporated to dryness and the residues obtainedused for isolating the fermentation products. The crude residue wasdissolved in methylene chloride and chromatographed through a 150 g.Florisil (synthetic magnesium silicate) column packed wet withSkellysolve B (hexanes) and eluted with 400 ml. fractions by gradientelution between l. of 4 percent acetoneSkellysolve B and 5 l. of 12percent acetone- Skellysolve B. Fractions 4 through 7 gave well formedcrystals. These fractions were combined and recrystallized from methanolwith Darco (activated charcoal) treatment to give 0.4 5 g. of7a-methylestrone (l l melting at 237 to 238 C.; this. 280 mu; =2,100.

Anal. Calcd. for C H O C, 80.21; H, 8.51. Found: C,

80.10;H, 8.34. I A small amount of 7a-methylestradiol can be isolatedfrom the more polar fractions.

EXAMPLE 2 7a-methylestrone (l l) A sterile medium was preparedcontaining 10 l. of tap water, 20 g. of comsteep liquors and 100 g. ofcommercial dextrose and adjusted to pH 5 with sodium hydroxide. This wasinoculated with 500 ml. of a vegetative growth of Sepiamyxa affim's(ATCC 6737) grown on thesame medium. The culture was stirred and aeratedat a rate of 0.1 l./minute. At the end of about 24 hours the pH was 7.To the fermentor, 2 g. of 7a-methyl-l9-nortestosterone (1) and 0.1 g. of3- ketobisnor-4-cholen-22-al. dissolved in 20 ml. of N,N-dimethylformamide was added. Aeration was continued for about 48 hours,the pH adjusted to 3 and the beer extracted four times with methylenechloride. Paper chromatography indicated the presence of7a-methylestrone l l) in the extract. The product (11) can be isolatedin accordance with the procedure described in example 1 to giveessentially pure 7amethylestrone (l 1).

Following the procedure of examples 1 and 2 but substituting7a-methyl-l9-nor-4-androstene-3,l7-dione for 7a-methyll9-nortestosteronel also yields 7a-methylestrone l l EXAMPLE 3 7a-methylestrone l 1 Amixture of 100 mg. of 7a-methyl-l9-nor-4-androstene- 3,17-dione, 40 mg.of 5 percent palladium on charcoal catalyst in 100 ml. of p-cymene(purified by passing through a column of alumina) was refluxed for about1.5 hours. The catalyst was removed by filtration and the filtrateconcentrated in a rotary evaporator. The residue was crystallized fromether to give 25 mg. of 7a-methylestrone l l), melting at 218 to 230 C.,xiilx. 279;:me; FZJSO. Infrared spectral analysis indicates it is thesmile compound as prepared in examples I and 2. The product can befurther purified by recrystallization or chromatdgraphy to giveessentially pure 'Ia-methylestrone l 1).

EXAMPLE 4 added. After standing for about 16 hours, methylene chloridewas added to the mixture and the DDQ filtered off. The filtrate wasdried and the solvent removed. The residue, weighing about 30 mg. wastriturated with methanol to give about 1 mg. of 7a-methylestrone (11)with xirlirx. 280 mu; e=2,350. Infrared analysis indicated that the thusproduced compound is the same as those prepared in examples 1, 2 and 3.

Following the procedure of example 4, but substituting selenium dioxideor tetrachloro-p-benzoquinone (chloranil) for DDQ, also yields7a-methylestrone l l EXAMPLE 5 7a-methylestrone l l) A pyrolysis tube 18inches l inches) equipped with thermocouples about 3 inches from eachend and packed with glass tubing cut in about V4 inch lengths was heatedtoabout 550 C. and heavy mineral oil passed through at a rate of 5ml.lminute until constant temperature of 500 to 510 C. at the top and540 to 560 C. at the bottom was obtained. A mixture of 10.4 g. of7a-methyl-l ,4-androstadiene-3, l 7-dione and 500 ml. of heavy mineraloil mixed in a Waring blender was added at a rate of 5 ml./minute,maintaining the above temperatures. The effluent was refrigerated forseveral hours and the crystalline precipitate collected, washedthoroughly with Skellysolve B anddried to give 6.7 g. of crude7amethylestrone (l 1). It was dissolved in hot methylene chloride,cooled and poured on a 350 g. Florisil column packed wet withSkellysolve B and eluted with 400 ml. fractions by gradient elutionbetween 5 l. of 4 percent acetone- Skellysolve B and 5 l. of 12 percentacetone-Skellysolve B. The desired product was contained in fractions 10through 21. The residues obtained from these fractions were combined andrecrystallized from methanol to give 3.75 g. of 70:- methylestrone l lhaving a melting point of 230 to 235 C.; [01],, (chloroform); xillfix.279 mu; r.==2,250. Nuclear magnetic resonance (NMR) spectra confirm theproposed structure and infrared spectral analysis shows it is the sameas the product obtained in examples 1, 2, 3 and 4.

EXAMPLE 6 7a-methylestrone l l) Treating7a-methyl-l,4-androstadiene-3,l7-dione in accordance with the proceduresdescribed in J. Amer. Chem. Soc. 86, 742, namely, with lithium anddiphenyl in the presence of diphenylmethane and employingtetrahydrofuran as solvent, yields 7a-methylestrone (1 l EXAMPLE 77a-methylestrone 3-cyclopentyl ether (1 l A mixture of 1 g. of7a-methylestrone (l) and l g. of cyclopentyl bromide is added slowly toa solution of sodium ethylate (prepared from 0.1 g. of sodium and 8 ml.of absolute ethanol). The reaction mixture is heated to reflux for about4 hours, the ethanol removed by distillation and the residue treatedwith a small amount of water. A precipitate of 7amethylestrone3-cyclopentyl ether (11) is obtained which is recrystallized from amixture of methylene chloride and methanol.

EXAMPLE 8 7a-methylestrone 3-methyl ether l 1) To 4 g. of7a-methylestrone (l l) in a solution containing 6.4 g. of potassiumhydroxide, 14 m1. of water and 21 ml. of methanol, 14 ml. ofdimethylsulfate was added dropwise with stirring and cooling sufficientto keep the reaction temperature at 25 to 35 C. At the same time asolution of 14 g. of potassium hydroxide in 28 ml. of water and 42 ml.of methanol was added dropwise at a rate that kept the pH of thereaction mixture above 10. The addition of the dimethylsulfate requiredabout 30 minutes and the addition of the potassium hydroxide solutionabout 1 hour. The reaction mixture was stirred an additional 1.5 hours.Water was added and the crystalline product collected on a filter,washed with water and dried to yield 3.8 g. of product (11) melting at155 to 163 C. It was recrystallized from methanol to give an analyticalsample of 7a-methylestrone B-methyl ether l 1) melting point 163 to 165C.; hrii zix. 276 mp; t=2,100; 286 mp4 F2,050.

Anal. Calcd. for C l-Q 2 C, 79.95; H, 9.39. Found: C,

Following the procedure of example 8 but substituting for dimethysulfatethe following:

1. diethylsulfate,

2. dipropylsulfate,

3. diisopropylsulfate,

4. dibutylsulfate,

5. di-s-butylsulfate,

6. dipentylsulfate,

7. dihexylsulfate,

8. diheptylsulfate and 9. dioctylsulfa'te, yields, respectively,

1. 7a-methylestrone 3-ethyl ether l l 2. 7a-methylestrone 3-propyl ether1 1),

3. 7a-methylestrone 3-isopropyl ether l l),

4. 7a-methylestrone 3-butyl ether l 1 5. 7a-methylestrone 3-s-butylether l 1),

6. 7a-methylestrone 3-pentyl ether l 1 7. 7a-methylestrone 3-hexyl etherl 1 8. 7a-methylestrone 3-heptyl ether l l and 9. 7a-methylestrone3-octyl ether l l EXAMPLE 9 7a-methylestrone 3-tetrahydropyranyl ether ll To a solution of 4. g. of 7a-methylestrone (l 1) in 40 m1. oftetrahydrofuran (purified by percolation through a column of alumina)and 8 ml. of freshly distilled dihydropyran,.0.4 ml. of phosphorusoxychloride was added dropwise with stirring under nitrogen and coolingin an ice bath. After the addition was completed the ice bath wasremoved and after an additional 15 minutes the reaction mixture waspoured into a mixture of saturated solution of sodium bicarbonate, etherand ice. The ether layer was separated, washed with dilute sodiumbicarbonate solution, water, dried over sodium sulfate and filtered. Thefiltrate was evaporated to dryness to give 5.1 g. of 7a-methylestrone3-tetrahydropyranyl ether l 1).

Following the procedure of example 9 but substituting for dihydropyranthe following:

l. dihydrofuran,

2. -hydroxymethyldihydropyran,

3. S-carboxydihydropyran, etc., yields, respectively,

1. 7a-methylestrone S-tetrahydrofuranyl ether l l 2. 7a-methylestrone3-(5-hydroxymethyl)pyranyl ether 3. 7a-methylestrone3-(5-carboxyl)pyranyl ether l l etc.

EXAMPLE l0 7a-methylestrone 3-acetate l 1) To 1 g. of 7a-methylestrone(l1), 2 ml. of pyridine and 1 ml. of acetic anhydride is added. Thereaction mixture is kept at room temperature for about 3 hours; water isthen added to precipitate the product (1 l) and destroy the excessacetic anhydride. Recrystallization from acetone and Skellysolve Byields 7a-methylestrone 3-acetate (l 1).

Following the procedure of example 10 but substituting for aceticanhydride the following:

1. benzoic acid anhydride,

2. propionic anhydride,

3. i-butyryl chloride,

4. valeryl chloride,

5. decanoyl chloride,

6. hexanoic anhydride,

7. sec. octanoic anhydride 8. capric anhydride,

9. undecyl anhydride,

l0. dodecanoyl chloride, etc., yields, respectively,

1. 7a-methylestrone 3-benzoate l 1),

7a-methylestrone 3-propionate l l), 7a-methylestrone 3-i-butyrate 1 l),7a-methylestrone 3-valerate l 1), 7a-methylestrone 3-decanoate (l l), 6.7a-methylestrone 3-hexanoate l l), 7. 7a-methylestrone 3-sec. octanoate(1 l (II-huh) 8. 7u-methylestrone 3-caproate (1 l), 9. 7amethylestrone3-undecanoate (l 1), l0. 7a-methylestrone 3-dodecanoate (l l etc.

EXAMPLE 1 l 7a-methylestrone 3-trimethylsilyl ether 1 1) To a suspensionof 4 g. of 7a-methylestrone (l) in 10 ml. of dry acetone, 6.5 ml. ofhexamethyldisilazane was added. The mixture was stirred for about 4 daysand then evaporated to dryness. The residue was dissolved in a mixtureof methylene chloride and Skellysolve B and chromatographed over a 250g. column of Florisil. Gradient elution between 5 l. of Skellysolve Band 5 l. of 10 percent acetonepercent Skellysolve B yielded 2.5 g. of7a-methylestrone-3-trimethylsilyl ether l l having a melting point of103 to 107 C.

Following the procedure of example 1 l but substituting otherdisilazanes for hexamethyldisilazane, such as symmetricaldiphenyltetramethyldisilazane, hexaamyldisilazane, etc., yieldsrespectively, 7a-methylestrone 3-phenyldimethylsi1yl ether l 1),7a-methylestrone 3-triamy1silyl ether l l etc.

The reactions of example 11 and the paragraph thereafter are preferablycarried out with the addition of a few drops of 2 ml. of trimethylsilylchloride.

In place of acetone in example 1 1, other inert dry solvents, such astetrahydrofuran, dioxane, methylene chloride and the like, can beutilized.

The 7a-methylestrone 3-silyl ethers (11) exhibit strong estrogenic,antifertility and gonadatropin suppressing activities. They can beadministered orally or, preferably, by injection.

As indicated above, the compounds of this invention are useful for theirestrogenic activity. Administration to mammals depends on the particularcompound involved, route of administration, severity of the conditionbeing treated and the individuals response thereto. In general, a doseof between about 0.01 mg. to about 5 mg. of each of the compoundsexemplified in examples 1 through 11 and embraced within formula 11 isgiven orally once a day, or subcutaneously or intramuscularly in a doseof 0.05 to 10 mg. weekly to monthly, in the treatment of conditionsincident to the foregoing activity when incorporated in conventionalpharmaceutical compositions.

The following examples illustrate the incorporation of the activeingredients of this invention in pharmaceutical formulation for use asestrogenics or antifertility agents.

EXAMPLE l2 Compressed tablets A lot of 10,000 compressed tablets, eachcontaining 0.05 mg. of 7a-methylestrone or 7a-methylestrone B-methylether is prepared from the following ingredients:

7a-methylestrone or 7a-methylestrone 3-methyl ether 0.5 g. Dicalciumphosphate 2,500 g. Methylcellulose, USP l5 cps.) 65 g. Talc, bolted 4503. Calcium stearate, fine powder 35 g.

passed through a No. 12 screen, mixed thoroughly with the talc andstearate and compressed into tablets.

EXAMPLE 13 Hard gelatin capsules A lot of 1,000 hard gelatin capsules,each containing 0.5 mg. of 7a-methylestrone or 7a-methylestrone 3-methylether is prepared from the following ingredients:

7a-methylestrone or 7a-methylestrone 3-rnethyl ether g. Lactose I50 3.Calcium stearate 2 g. Talc 3 g.

The lactose, talc and stearate are mixed well and incorporated into themixture. The whole is mixed well and filled into two-piece hard gelatincapsules.

EXAMPLE 14 Soft gelatin capsules A batch of 1,000 soft gelatin capsules,each containing 0.5 mg. of 'la-metylestrone or 7a-methylestrone 3-methylether and corn oil is prepared from the following materials:

7a-methylestrone or 7a-merhylestrone 3-methyl ether Corn oil A uniformdispersion of the active ingredient in the corn oil is prepared and thedispersion filled into soft gelatin capsules by conventional means.

EXAMPLE l5 Aqueous oral suspension An aqueous oral suspension containingin each 5 ml. 0.5 mg. of 7a-methylestrone or 7a-methylestrone 3-methylether is prepared from the following materials:

7a-methylestrone or 7a-methylestr0ne EXAMPLE l6 Aqueous suspension forinjection A suspending vehicle is prepared from the following materials:

Polyethylene glycol 4000 30 g. Potassium chloride l l.2 g. Polysorbate80 2 g. Methylparaben L8 g. Propylparaben 0.2 g. Water for injectionq.s. I000 ml.

The parabens are added to a major portion of the water and are dissolvedtherein by stirring and heating to 65 C. The resulting solution iscooled to room temperature and the remainder of the ingredients areadded and dissolved. The balance of the water to make up the requiredvolume is then added and the solution sterilized by filtration. Thesterile vehicle thus prepared is then mixed with 0.5 g. of7a-methylestrone or 7a-methylestrone 3-methyl ether which has beenpreviously reduced to a particle size less than about 10 microns andsterilized with ethylene oxide gas. The mixture is passed through asterilized colloid mill and filled under aseptic conditions into sterilecontainers which are then sealed.

Each milliliter of this suspension contains 0.5 mg. methylestrone or7a-methylestrone 3-methyl ether.

As indicated above, the compounds of this invention, in addition totheir use as estrogenics and antifertility agents, when combined withprogestins, e.g., 6a-methyl-l7a-hydroxyprogesterone l7-acetate(Provera), 7a-methyl-l7a-ethynyll9-nortestosterone,l7a-hydroxy-6-methyll 6-methylene-4,6- pregnadiene-3,20-dione l7-acetate(Melengestrol acetate), etc., are useful for the prevention of ovulationin mammals. Administration to mammals depends on the particularprogestin and estrogen involved and the individual s response thereto.In general, a dose of between about 0.01 mg. to about 5 mg. of each ofthe estrogens exemplified in example 1 through ll and embraced withinformula ll plus between about 1 mg. to about mg. of a progestin is givenat such time(s) in the mammalian ovulatory cycle as is suitable for theprevention of ovulation. v

The following examples illustrate the incorporation of the activeingredients of this invention with progestins in pharmaceuticalformulation for use as anovulatory agents.

EXAMPLE I 7 ba-methyl-l'la-hydroxyprogestcrone l7-acetate 5 oz.7a-methylestrone or 7a-methylcstrone B-methyl ether 23 grains Lactose 3lbs.

The finely powdered active ingredients and lactose are mixed well andgranulated with syrup-starch paste. Starch and calcium stearate are usedas lubricants in the compressing step.

EXAMPLE 1 8 Oral tablets 10,000 tablets for oral administration areprepared from the following types and amounts of ingredients. Eachtablet contains 10 mg. of 6a-methyl-l7a-hydroxyprogesterone 17- acetateand 0.05 mg. of 7a-methylestrone or 7a-methylestrone 3-methyl ether.

6a-methyll 7a-hydroxyprogestcrone l7-ucetate I00 g. 7u-methyleslrone or7a-methyl estrone J-methyl ether 0.5 g. Lactose 2,600 g.

The finely powdered active ingredients and lactose are mixed well andgranulated with syrup-starch paste. Starch talc and calcium stearate areused as lubricants in the compressing step.

EXAMPLE 19 Oral aqueous suspension An aqueous suspension for oraladministration, containing in each teaspoonful (approximately 5 ml. 5mg. of 6a-methyl l7a-hydroxyprogesterone l7-acetate and 0.2 mg. of 7a:-methylestrone or 7a-methylestrone 3-methyl ether is prepared from thefollowing types and amounts of ingredients:

a-methyll 7a-hydroxyprogesterone l7-acetate l g. 7a-methyleslrone or7a-methylestrone S-methyl ether 40 mg. Preservative 2 g. Flavor, q.s.

Purified water USP, a.s. and I000 rnl.

The preservative and flavor are dissolved in the water. The micronizedactive ingredients are added and the whole is homogenized.

EXAMPLE. 20

Oral gelatin capsules One thousand gelatin capsules for oraladministration, each containing 10 mg. of6a-methyl-l7a-hydroxyprogesterone l7- acetate and 0.05 mg. of7a-methylestrone or 7a-methylestrone 3-methyl ether are prepared-fromthe following types and amounts of materials:

6a-methyll 7a-hydroxyprogesterone l'l-acetlte g. 7u-methylestrone or7a-methylestrone B-rnethyl ether 50 mg.

Ingestible oil. q.a.

The rnicronized active ingredients and the oil are mixed and the mix isencapsulated by the usual techniques into gelatin capsules.

EXAMPLE 21 Oral tablets Following the procedure of example 17, 5,000tablets are prepared from the following types and amounts ofingredients. Each tablet:

5 mgv 6cr-methyll 7a-hydroxyprogesterone l7-acetate 25 g. 0.0l rng.'Ia-methylestrone or 7a-rnethylestrone 3- methyl ether 50 mg. I50 mg.lactose 750 g. 3 mg. acacia g. 65 mg. starch, bolted 325 g. 3 mg.calcium stearate I5 g.

Tablets equally suited for the inhibition of ovulation are prepared byusing 250 and 1000 mg., respectively, of the 7amethylestrone or7q-methylestrone 3-methyl ether in place of the 50 mg. in the aboveformulation.

While the procedures described above in examples 17 through 21 recitethe use of the progestin 6a-methyl-l7ahydroxyprogesterone 17-acetate,other progestational compounds can be substituted therefor to providesimilarly effective anovulatory pharmaceutical formulations; e.g.,7amethyl- 1 7a-ethynyll 9-nortestosterone, l 7a-hydroxy-6- methyl- 16-methylene-4,6-pregnadiene-3,20-dione l7-acetate, l7-hydroxyl 9-norl7a-pregn-5( l0)-en-20-yn-3-one, l9-norl7a-pregn-4-en-20-yne-3B, l7-diol3 ,1 7-diacetate, l7- hydroxyl 9-norl 7a-pregn-4-en-20-yn-3-onel7-acetate, 601,2 1 -dimethyll 7B-hydroxy-4-pregnen-20-yn-3-one, 6-

chlorol 7a-hydroxy-4,6-pregnadiene-3,ZO-dione l 7-acetate. etc., can beemployed instead of 6a-methyl-l7a-hydroxyprogesterone l7-acetate.

In place of 7a-methylestrone (ll) and 7a-methylestrone 3- methyl ether ll other ethers and the acylates of formula I I, such as 7a-methylestrone3-cyclopentyl ether (l 1), 7amethylestrone 3-tetrahydropyranyl ether ll), 7a-methylestrone S-trimethylsilyl ether (l l), 7a-methylestrone3-acetate (11), etc., can be employed.

We claim:

1. An oral pharmaceutical composition comprising:

a. about 0.01 to about S-mg. of a compound of the formula wherein R isselected from the group consisting of hydrogen and methyl, and

b. about 1 to about 100 mg. of a progestin, dispersed in apharmaceutical carrier. 2. A composition in accordance with claim 1wherein the progestin is 7a-methyll 7a-ethynyll 9-nortestosterone.

3. A method of preventing ovulation in ovulating mammals wherein R isselected from the group consisting of hydrogen and methyl, and

b. a progestin.

4. A method of preventing ovulation in ovulating mammals in accordancewith claim 3 wherein the progestin is 70:- methyll 7a-ethynll 9

2. A composition in accordance with claim 1 wherein the progestin is 7Alpha -methyl-17 Alpha -ethynyl-19-nortestosterone.
 3. A method ofpreventing ovulation in ovulating mammals comprising: orallyadministering to ovulating mammals an effective amount of a. a compoundof the formula
 4. A method of preventing ovulation in ovulating mammalsin accordance with claim 3 wherein the progestin is 7 Alpha -methyl-17Alpha -ethynyl-19-nortestosterone.